Q1.

Multiple Choice: select the best possible answer: (14 point)

1. Some parameters will be altered during the Lag phase:

a. Increase of nutrient supply.

b. Increase in inhibitors.

c. Change in pH.

d. a and c.

2. The Term X in the below equation indicates: dX/dr = uN

a. The biomass.

b. The mass of cells in grams.

c. A measure of growth for filamentous microorganisms.

d. All of the above are correct.

3. Catabolite repression is best defined as:

a. Inhibition of microbial growth.

b. Inhibition of the targeted product by the substrate.

c. Microorganisms used in industry can't metabolize some carbon sources.

d. Inhibition of the targeted product by the end product.

4. In the below equation, which parameter is identified as the saturation constant:

a. µ

b. µ max

c. ks substrate con when growth rate = ½ maximum growth rate

d. S

5. Recombinant DNA techniques are powerful tools for strain development via:

a. Synthesize a new bacterium that contains powerful RNAs.

b. cloning a DNA fragment into a bacterium which adds in required genes

c. produce rRNA

d. none of the above

6. Balanced growth is achieved when:

a. The growth rate reaches its lowest value

b. The minimal growth nutrient is utilized

c. Every extensive component of the growing system increases by the same factor

d. Glucose is added at very limiting concentration

7. The concept of using a deregulated mutant in biotechnology is to overcome the problem of:

a. The problem of mixed products.

b. The problem of catabolite repression.

c. The problem of undesirable mutation.

d. The problem of feedback inhibition.

8. The following is used in industrial fermentation to produce food and drinks

a. Yeast

b. Algae

c. Vitamines

d. Vaccines

9. Suitable host in genetic engineering to introduce DNA fragments of donor is

a. Yeast

b. Bacillus subtilis

c. Escherichia coli

d. Bacteriophages

10. The donor DNA fragments and vector DNA fragments are joined together by:

a. DNA ligase

b. Plasmid

c. Auxin

d. Cytokinin
11. Yield coefficient represents

a. total biomass or product produced

b. conversion efficiency of a substrate into product

c. conversion rate of a substrate into biomass or product

d. production time of biomass or product

12. The lowest biomass yield in a culture of Escherichia coli will be in

a. an aerated batch culture containing a initial high concentration of glucose

b. an aerated batch reactor containing an initial low concentration of glucose

c. an aerated fed-batch reactor having a low glucose concentration

d. an aerated continuous reactor having a low glucose concentration

1. Four Arab countries (- &-&-&-) were conducting GM trials, but only one country (- ) released GM
maize (2008)

a. Egypt, Oman, Morocco & Syria/Egypt

b.Egypt, Jordan, Tunisia & Syria Oman

c. Egypt, Tunisia, Morocco & Sudan/Egypt

d. Egypt, Jordan, Tunisia & Syria/Jordan

2. After the discovery of restriction enzyme, the following processes were known

a. Hybridoma techniques and tPA plasminogen

b. Hormone production, herbicide tolerance and viral vaccination

c. Antibiotic production and fermentation

d. Immobilization of antibiotics and hazardous waste management

3. Biotechnology is the art of science that makes use of:

a. Cells and cell products

b. Engineering principles

c. Molecular biological and cell sciences

d. All of the above are correct

4. Strain development is a principal step in industrial biotechnology which includes

a. Modification of culture conditions

b. Mutation of the wild type

c. Optimization of the fermentation conditions

d. All of the above are correct

5. All of the following are biotechnological products produced via the five main processing steps
except:

a. Enzymes

b. Antibiotics

c. Stem cells

d. None of the above

6. One of the following is not considered an outcome of biotechnology operations:

a. Human insulin

b. Herbicide resistant plants

c. Reduction of the global death rate

d. Production of GM food

7. In the post recombinant DNA era, a human therapeutics were developed:

a. Insulin, hGH, viral vaccine

b. Insulin, penicillin, hGH

c. tPA, hGH, penicillin

d. All of the above tpa

8. Plasminogen activation (tPA) is responsible for:

a. Enhancing the formation of blood clot.

b. Breaking down blood clot

c. Regulate blood hormones

d. Regulate blood pressure

9. Oxychemicals (Glycerol, acetic acid, Acetone) can be produced by:

a. Fermentation of hydrocarbon

b. Hydrolysis of lignocellulosic material

c. Fermentation of sugar

d. b& c

10. In biotechnology, "process engineering" is a process which includes:

a. Fermentation and bacterial growth

b. Fermentation and genetic engineering

c. Fermentation and isolation of product

d. All of the above.

11. upstream processing (USP) includes:

a. strain improvement and optimization of growth condition.

b. processing of end product

c. strain improvement or processing of c-source

d. all of the above.

12. if activation of an inducible enzyme is needed, then the following should be added to the
reaction medium:

a. substrate

b. inducer

c. raw material

d. all of the above

13.methyl nitrosoguanidine (NTG) is:

a. a mutagen

b. an inducer

c. an inhibitor

d. used to cause deamination

14. UV is used to:

a. Induce transition mutation

b. Cause dimerization of thymine

c. Cause chromosomal mutation

d a& b

15. Guidelines of the raw materials include:

a. Nature of the raw material

b. The targeted end product

c. Microorganisms used

d. All of the above

16. optic development through mutation or cloning is an important step in biotechnology

operations. Which of the following processes is responsible for achieving that goal:

a. Downstream processing

b. Upstream processing

c. Fermentation

d. Pure product process

18. optimization of the upstream processing requires:

a. strain improvement

b. optimizing growth condition

c. processing of the raw materials

d. all of the above

19. microorganism used in industrial fermentation could be any one of the following except:

a. bacteria

b. actinomycetes

c. fungi

d. macrophages
20. whyion of mutant after exposing the bacteria to mutagens depends on:

a. Type of bacteria

B Desired mutation

C. Resistance to chemicals

d. Temperature sensitivity

21. 5-bromouracil is a mutagen that causes:

a. Transition mutation, A to T

b. Point mutation, replaces A to G

c. Point mutation, replaces C to T

d. None of the above

24. Plasmids are:

a. Cloning vectors naturally occurring in prokaryotic cells

b. Chromosomal DNA that is important of the cell life

c. Circular DNA fragments linked to the genome of the cell

d. None of the above are correct

5. In the Precombinant era,

a. Penicillin was produced

b. Monoclonal antibodies were purified

c. Growth hormone was discovered

d. Vaccines was discovered

6. Hybridoma technique is the technique which is related to

A. Production of Antibiotics

b. Production of monoclonal antibodies

C. Production of hydrophobic molecules

d. All of the above

Q2. Guess which of the following statements are True or False? (10 points)

• It is good to adjust the flow rate to be slightly lower than Dm. F

• In the continuous culture under steady state condition, u=D. T
• It is difficult to reduce the time at which the bacteria spend in the Lag Phase. F
• Growth rate of cells during the Log phase remains constant. T
• The bacterial cells will be doubled during the Lag phase in the batch culture. F
• Flocculation process is particularly used in production of enzymes. F
• Rolling circle is a replication mechanism for the DNA phage which form concatemer. T
• In biotechnology, sometimes it is important to deal with aggregated cells; which simply can be
done by heat. T
• At low flow rate X decreases and may reach to 0. F
• Packed bed reactor is used for the production of enzymes and small molecule bioconversion. F
• The Monod equation is not applicable when intra-substrate concentration is reduced during fast
growth. T
• The wash-out Dm will never equal the maximum specific growth rate µm. F
• It is difficult to reduce the time at which the bacteria spend in the Lag. Phase. F
• Growth rate of cells during the Log phase remains constant. T
• The bacterial cells will be doubled during the Lag phase in the batch culture. F
• Flocculation process is particularly used in production of enzymes. F
• Rolling circle is a replication mechanism for the DNA phage which form concatemer. T
• Performing the Gram stain (G-, G+) for bacteria is important before determining the proper
disintegration technique. T
• For collecting microbial cells in SCP, first they should be heated to be aggregated followed by
centrifugation. T
• During the filtration process of antibiotics, filtration rate will depend on viscosity, type (nature)
of cells, and acidity. F

Q3. Answer the following questions: (30 points)

1. Describe in general terms (e.g., no need for media formulations) a procedure for growing a
proteolytic bacterium and enhancing the yield of the proteolytic enzyme. (6 points)

But the bacteria in complete medium ( nutrient broth or yeast extract) and but them in the shaker
for 24h at 34-40C, make a serial dilution and chose 2 dilutions and do plating on NA and let them
grow and see the clear zone the one that has more clear zone is the highest activity. And we should
optimize the condition by providing all the requirements for the culture

2. What are the factors that affect the filtration process? (3 points)

Viscosity, pore size, cost, reuse of the filter, particle selectivity, presence of antifoam, dead volume,
sterilization of the system.
 3. What is specific growth rate (u), What are the factors that control it? (4 points)

A measurement of how fast the particular bacterium grows in a particular environment. The factor is
the substrate, substrate-specific constant, maximum growth rate.

4. Compare between the two major types of filters (Depth and Absolute filter). (4 points)
Items Depth filter Absolute filter
Nature Normal filter paper, glass wool, asbestos Membrane filter (small pore size 0.25-
0.45 Mm)
purpose Fungal mycelium Used in bacterial culture to separate cell
from the medium

5. What are the differences between a batch culture and continuous culture. (5. points)
items Batch culture Continuous culture Fed-batch
System Closed system process Open system Begin with a small amount
of nutrient and gradually
increase it (catabolic
repression)
Addition of food No addition during the process Added during the process Added during the process

Removing of Not removed Removed continuously

wastes
Growth phase Lag, log, stationary, death Lag, log, stationary, death

When used Formation of amino acid, Ethanol production, Production of penicillin

enzyme. wastewater treatment,
Decomposition ( sewage SCP production
treatment, bioremediation)
Antibiotics

Ph, nutrient con, Not constant Constant

metabolic product

6. Monod equation is not always applicable to continuous fermentation State the condition(s) at
which Monod equation is not applicable? (2 points)
When the concentration of substrate inside the cell is lower than outside the cell because of fast
growth.
7. Explain why It is preferable to use a recombinant phage production of E. coli DNA ligase. (2
points)
Because the DNA ligase gene induces five-hundredfold overproduction of the enzyme.
8. What is a bioreactor (2 points)
an apparatus in which a biological reaction or process is carried out, especially on an
industrial scale.
Q2 Answer the following questions: (22 points) 1)

1. What is the role of environmental biotechnology? (3 points)

Bioremediation use microorganisms to break down some chemicals, biomining is use

microorganisms to extract the minerals from the rocks, used as biosensor which reacts with
chemicals that will act as an indicator for these chemicals.

2. If you run an enrichment experiment to screen for bacteria producing protease enzymes. What
would you do after completing the enrichment experiment? (2 points)

-select the most potent strain which produces protease ( high enzyme activity) and preserve the
other strain to act as alternatives in the future.

3. In case of using lignocellulosic material as a raw material for bacterial growth, what is the first
step you have to do? How? (3 points)

Make a pre-treatment by letting some organisms to utilize the material and by doing hydrolysis be
chemical or enzymatic.

4. How would you enrich for auxotrophic mutant? How would you recognize an auxotrophic
mutation? (4)

Enriching by the bacteria in complete medium and for the mutant recognition we take a replica
platting of the plate and but it in minimal medium and see the bacteria that grow is have the
mutation.

5. Give two examples of biofertilizers resulted from biotechnology? Why did you consider them as
biotechnological product? (3 points)

Algae, Rhizobium, because these organisms help the plant in many ways for example
nitrogen fixation

6. Why it is necessary to improve the strain used in fermentation?

To optimize the growth condition, to adapt growth on the RM, to determine the growth
characteristics, to increase the yield.

7. Although remarkable advances in biotechnology was achieved, there are some public concerns
which hold some people from using the biological products. List the concerns:

Vaccines: Administration of live GE microorganisms.  Protection of plants against pests involves the
introduction of bacteria in or within the plants.  Establishment of new symbiotic relations. Harm
to nontarget species. Disruptive effects on biotic communities.

8. Why the microorganisms use the raw material? List the criteria which are used to select the raw
material?

The raw material is used as a source of carbon that is needed for ATP production. Criteria: the target
end product, nature of the raw material, condition of the process, microorganisms used.
9. What is biopharming? Give example

Is the use of agriculture plants for the production of a useful molecule for non-food, feed pr fibre
application.

Explain why: (8 points)

a. Preservation of excellent strains is important in biotechnology

For the future as an alternative source because some gene have regulation mechanisms ( off/on)

b. The 5 ends ds lambda genome exhibit ss regions of 12 nucleotides

To produce cos site and cohesive termini that allow the linear DNA to be circulated during the
injection, so the 5 ends produce this end to stick and become circular.

c. Food preservatives sometimes considered as biotechnological product.

Add some bacteria such as acetic acid bacteria which produce lactic acid which kills other types of
bacteria.

d. Seek and discard principle is applied in selecting microorganism

To select for the best producing strain to have a high production and for strain improvement

e. During cloning with plasmid the vector and the forging DNA are cleaved with the same enzyme.

To get a sticky end and both become complementary to each other to stick to each other.

f. Culture media containing glucose must be carefully sterilized to avoid:

To avoid Millard reaction because some time the amino acid reacts with the reducing sugar to give
brown colour glycosylamine which is toxic for bacteria
 Stirred-tank reactor Bubble column Airlift Packed bed ( fixed bed)
-widely used - well-organized sparger -consist of 2 pipes -It consists of a tubular
-Fungal mycelia nozzles to produce the connected at the top and column packed with the
-production of biopolymers same bubble size and bottom. support that retains (or
- used if the product has well mixing. - the introduction of the entraps) microbial and
high viscosity. air in the pipes resulting fungal cells.
in the circulation of the - They are commonly used
reactor content between for small molecule
the risers and the bioconversions.
downcomer
-The circulation is
induced by the difference
in density between the
aerated liquid in the riser
and the liquid in the
down comer.
FLOCCULATION Flotation Filtration Centrifugation
-used for very small -Cells adsorbed on the - depth filter -depend on sedimentation
molecule gas bubbles and floated -absolute filter rate.
- treatment SCP and as foam. Factors affect the
sewage treatment sedimentation rate:
-flocculation agent: Diameter of particle,
inorganic salt, organic density of the particle and
polyelectrolyte, mineral liquid, radius of the
hydrocolloid. centrifugation head,
angular velocity and
dynamic viscosity

Gel filtration Adsorption Ion exchange Affinity chromatography

chromatography chromatography
- The gel filtration - Adsorption separates ions and -method of separating
chromatography Chromatography polar molecules a biomolecule from a
is based on the involves the separation based on their affinity mixture, based on a
molecular size and the of a chemical mixture to the ion exchanger highly specific
hydrodynamic volume based on the macromolecular binding
of the components interaction of the interaction between the
solvent biomolecule and beads

Isoelectric focusing

Separation based on ph
 Solvent extraction Aqueous extraction
•Organic solvents Applied for organic solvent labile materials
•Solvent is immiscible(not mixed) with water • A salt with hydrophilic polymers is used
•Ideal transfer of the target product • Water is the main solvent in both phases
•The target product will transfer quantitatively • Cells remain in one of the phases
to the solvent phase • The enzyme transfers to the other phase
•Applied to antibiotics such as penicillin using without any loss of activity.
amyl or butyl acetate • Example of products is human fibroblast
interferon

- If you discover a new enzyme what should you do : GRAS

Description of the enzyme and its source, The method of enzyme preparation:
• Complete composition of the medium
• Equipment used in production
• Fermentation conditions: aeration, T, pH & agitation
• Toxicological of the enzyme or the microorganisms : nonpathogen organisms, not produce toxins
or antibiotics

Steps of starch processing:

Liquefaction Saccharification Isomerization

Type of • Using  -amylase from B. - a-1,4 glucoamylase

enzyme used licheniformis from A. niger - highly purified glucoamylase
Convert starch to low DE - Pullulanase from from S. rubiginosus
maltodextose Bacillus sp.
Ph • Near neutral pH Mild acidic pH pH 7.6

Temperature 95 – 107 C Low temp 55-57 C

Enzyme Stabilized by Ca+2 ions . Mg+2 (1.5mM) for activity, Ca+2 ,

needed ions bisulfite added for preservation and
also to stabilize mg and enzyme
DE 5 - 15 97 The product is 42 DI HFCS.

Immobilized to granular solid Recovery • Filtration • The enzyme can be desorbed,

carrier: DEAE diethyl amine Decolorization • Pure and purified and re-immobilized again for
Ethyl Cellulose (30%)+Food crystalline glucose further production.
grade polystyrene
50%+Titanium dioxide (20%)
(electrostatic interaction).
What is the importance of bioinformatics if you have a sequence?